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基于SSR标记的山西省油松山脉地理种群遗传结构与地理系统

武文斌 贺快快 狄皓 钮世辉 马彦光 张子杰 李悦

武文斌, 贺快快, 狄皓, 钮世辉, 马彦光, 张子杰, 李悦. 基于SSR标记的山西省油松山脉地理种群遗传结构与地理系统[J]. 北京林业大学学报, 2018, 40(10): 51-59. doi: 10.13332/j.1000-1522.20180057
引用本文: 武文斌, 贺快快, 狄皓, 钮世辉, 马彦光, 张子杰, 李悦. 基于SSR标记的山西省油松山脉地理种群遗传结构与地理系统[J]. 北京林业大学学报, 2018, 40(10): 51-59. doi: 10.13332/j.1000-1522.20180057
Wu Wenbin, He Kuaikuai, Di Hao, Niu Shihui, Ma Yanguang, Zhang Zijie, Li Yue. Genetic structure and geographic system of geographical population of Pinus tabuliformis mountain range based on SSR in Shanxi Province of northern China[J]. Journal of Beijing Forestry University, 2018, 40(10): 51-59. doi: 10.13332/j.1000-1522.20180057
Citation: Wu Wenbin, He Kuaikuai, Di Hao, Niu Shihui, Ma Yanguang, Zhang Zijie, Li Yue. Genetic structure and geographic system of geographical population of Pinus tabuliformis mountain range based on SSR in Shanxi Province of northern China[J]. Journal of Beijing Forestry University, 2018, 40(10): 51-59. doi: 10.13332/j.1000-1522.20180057

基于SSR标记的山西省油松山脉地理种群遗传结构与地理系统

doi: 10.13332/j.1000-1522.20180057
基金项目: 

中央高校基本科研业务费专项 2015ZCQ-SW-02

“十三五”国家重点研究计划 2017YFD0600500

详细信息
    作者简介:

    武文斌。主要研究方向:森林遗传学与针叶树遗传改良。Email:wwb474802665@163.com 地址:100083  北京市海淀区清华东路35号北京林业大学生物科学与技术学院

    责任作者:

    李悦,博士,教授。主要研究方向:森林遗传学与针叶树遗传改良。Email:liyue@bjfu.edu.cn 地址:同上

  • 中图分类号: S791.254

Genetic structure and geographic system of geographical population of Pinus tabuliformis mountain range based on SSR in Shanxi Province of northern China

  • 摘要: 目的分析油松种源区的遗传结构能够为油松起源探索和引种地人工林种群溯源等提供理论依据。方法本文利用SSR分子标记分析了山西油松5个山脉地理种群的遗传结构和地理系统。试验方法分别用了8对多态性较高引物和4对卡方检测差异显著性引物进行分析。结果结果显示:关帝山遗传变异最丰富,中条山遗传变异最低;8对引物分析显示5个山脉地理种群分化较小(FST=0.0328),4对引物分析显示5个山脉种群存在中等程度分化(FST=0.0519);5个山脉种群遗传距离和地理距离存在显著相关性;遗传多样性参数与1月均温有负相关趋势,年均降水与1月均温比值和遗传多样性参数有正相关趋势。结论研究发现山西省主要山脉油松地理种群间有相对较小的遗传分化和种群内有丰富的遗传多样性;SSR标记可以有效显示各种群的遗传结构特点,而筛选得到的有限差异显著标记能更有效的用于油松的种群遗传分析;生境气象因子的水热相对水平对种群遗传多样性有主导影响,相对较高的水热比利于种群保持较高的遗传多样性。

     

  • 图  1  8对引物(左)/4对引物(右)分析油松种群的遗传距离UPGMA聚类图

    Figure  1.  Eight pairs of primers (left)/four pairs of primers (right) for analyzing genetic distance among populations of P. tabuliformis in Shanxi Province by UPGMA clustering

    表  1  供试油松山脉种群地理位置、气候条件和样本数

    Table  1.   Geographical location, climatic conditions and sample number of the Pinus tabuliformis mountain range populations to be tested

    山脉
    Mountain range
    经度范围
    Longitude range
    纬度范围
    Latitude range
    年均温
    Annual mean temperature/℃
    1月均温
    Average temperature in January/℃
    7月均温
    Average temperature in July/℃
    年均降雨量
    Annual mean precipitation/mm
    样本数/林分数
    Sample number/stand number
    GCS 111°15′~112°33′E 38°10′~38°55′N 7.3 -8.7 21.5 447.6 52/6
    GDS 110°40′~111°50′E 36°48′~37°42′N 8.4 -7.2 22.0 489.6 66/6
    THS 113°24′~113°41′E 35°48′~38°47′N 7.1 -7.6 20.2 509.9 32/3
    TYS 112°01′~112°06′E 36°37′~38°36′N 10.5 -5.3 24.2 402.5 20/2
    ZTS 111°53′~112°08′E 35°32′~35°55′N 10.5 -3.4 23.2 567.8 18/2
    注:GCS代表管涔山,GDS代表关帝山,THS代表太行山,TYS代表太岳山,ZTS代表中条山。下同。Notes: GCS stands for Guancen Mountain, GDS stands for Guandi Mountain, THS stands for Taihang Mountain, TYS stands for Taiyue Mountain, ZTS stands for Zhongtiao Mountain. The same below.
    下载: 导出CSV

    表  2  油松PCR检测SSR引物

    Table  2.   SSR primers for PCR detection of P. tabuliformis

    引物名称
    Primer name
    前引物序列
    Forward primer sequence(5′→3′)
    后引物序列
    Back primer sequence(5′→3′)
    荧光修饰
    Fluorescent modification
    片段长度
    Fragment length/bp
    J9 GTTTGCAGTGAAAGCATGAAAG GCACCAATTCCTTCTCAAATTC HEX 244~250
    J10 GTCGACACTCCAGGGTAGATTC ATATCATCAGCTAATTGTGCGG TAMRA 254~257
    J12 TATGCATGTAACGGTAGCCTTG GCAATTGTTCTATGGTCAGGGT ROX 471~477
    J20 CACCTCCGTAGTTTGATGTTCC CGATGTATCGTGTACACAGCCT FAM 150~171
    J29 AGTCCGAATGTCTTCTTTCTGC TATGGAACGAATCAGAGATGACG FAM 191~200
    J42 AACCTGTCATCCAGTTCCTGTT TTGTCAAATTCCAATTCAGCAC TAMRA 251~269
    J48 GAAGAGGAAGACGAAATGGATG CTTTACATTTACCGCCTCTGCT ROX 262~268
    J50 TCATCCATTTCAATAGCACGAC GTAGCTGCTTGGCCTGATTATC HEX 235~244
    下载: 导出CSV

    表  3  山西5山脉油松种群各位点等位标记频率

    Table  3.   Frequency of the mark in five mountain populations of P. tabliformis in Shanxi Province

    标记Marker 群体Population 标记Marker 群体Population
    位点
    Loci
    等位标记
    Allele
    GCS GDS THS TYS ZTS 位点
    Loci
    等位标记
    Allele
    GCS GDS THS TYS ZTS
    J9 A 0.144 2 0.242 2 0.218 8 0.150 0 0.166 7 J42 A 0.038 5 0.045 5 0.171 9 0.025 0 0.027 8
    B 0.855 8 0.757 8 0.781 2 0.850 0 0.833 3 B 0.211 5 0.303 0 0.390 6 0.225 0 0.222 2
    J10 A 0.692 3 0.676 9 0.609 4 0.675 0 0.750 0 C 0.740 4 0.628 8 0.406 2 0.750 0 0.750 0
    B 0.307 7 0.323 1 0.390 6 0.325 0 0.250 0 D 0.000 0 0.022 7 0.000 0 0.000 0 0.000 0
    J12 A 0.823 5 0.882 8 0.937 5 0.875 0 1.000 0 E 0.009 6 0.000 0 0.015 6 0.000 0 0.000 0
    B 0.176 5 0.117 2 0.062 5 0.125 0 0.000 0 F 0.000 0 0.000 0 0.015 6 0.000 0 0.000 0
    J20 A 0.040 0 0.072 6 0.048 4 0.075 0 0.093 8 J48 A 0.200 0 0.269 2 0.312 5 0.289 5 0.027 8
    B 0.890 0 0.774 2 0.806 5 0.800 0 0.593 8 B 0.750 0 0.723 1 0.687 5 0.684 2 0.972 2
    C 0.070 0 0.137 1 0.129 0 0.075 0 0.312 5 C 0.050 0 0.007 7 0.000 0 0.026 3 0.000 0
    D 0.000 0 0.016 1 0.016 1 0.050 0 0.000 0 J50 A 0.000 0 0.015 2 0.000 0 0.000 0 0.000 0
    J29 A 0.009 6 0.015 2 0.000 0 0.000 0 0.000 0 B 0.125 0 0.159 1 0.187 5 0.175 0 0.166 7
    B 0.932 7 0.878 8 0.984 4 0.950 0 0.944 4 C 0.875 0 0.818 2 0.812 5 0.825 0 0.833 3
    C 0.048 1 0.098 5 0.000 0 0.050 0 0.055 6 D 0.000 0 0.007 6 0.000 0 0.000 0 0.000 0
    D 0.009 6 0.007 6 0.015 6 0.000 0 0.000 0
    下载: 导出CSV

    表  4  8个位点等位标记频率差异显著性χ2检测表

    Table  4.   Eight pairs of polymorphic primers for chi-square detection

    引物Primer J20 J9 J42 J12 J29 J50 J10 J48
    PP value 0.019* 0.344 0.002** 0.034* 0.452 0.857 0.676 0.007**
    注:*代表群体中等位标记频率差异显著(P < 0.05),**代表群体中等位标记频率差异极显著(P < 0.01)。Notes:* represents allelic frequency in populations is significantly different (P<0.05),** represents allelic frequency in populations is very significantly different(P < 0.01).
    下载: 导出CSV

    表  5  山西5山脉油松种群遗传多样性参数

    Table  5.   Genetic diversity parameters of five mountain populations of P. tabuliformis in Shanxi Province

    山脉Mountain range Na Ne I Ho He
    8对引物
    Eight pair primers
    4对引物
    Four pair primers
    8对引物
    Eight pair primers
    4对引物
    Four pair primers
    8对引物
    Eight pair primers
    4对引物
    Four pair primers
    8对引物
    Eight pair primers
    4对引物
    Four pair primers
    8对引物
    Eight pair primers
    4对引物
    Four pair primers
    GCS 2.7500 3.0000 1.4367 1.4992 0.5000 0.5733 0.3113 0.3198 0.2918 0.3264
    GDS 3.1250 3.2500 1.5825 1.6464 0.5970 0.6485 0.3722 0.3860 0.3559 0.3774
    THS 2.6250 3.2500 1.6442 1.8140 0.5536 0.6679 0.3876 0.4002 0.3407 0.3885
    TYS 2.5000 3.0000 1.4854 1.5621 0.5209 0.6129 0.3030 0.2809 0.3175 0.3588
    ZTS 2.1250 2.2500 1.4193 1.4670 0.4187 0.4178 0.2491 0.1788 0.2597 0.2531
    均值Mean 2.6250 2.9500 1.5136 1.5977 0.5180 0.5841 0.3246 0.3131 0.3131 0.3408
    注:Na.观测等位标记数;Ne.有效等位标记数;I. Shannon多样性指数;Ho.观测杂合度;He.期望杂合度。下同。Notes: Na, observed allelic marker number;Ne, effective allelic marker number;I, Shannon diversity index;Ho, observational heterozygosity;He, expected heterozygosity. The same below.
    下载: 导出CSV

    表  6  各位点在山西5山脉油松种群中的F统计量

    Table  6.   F-statistic for each locus in the five mountain populations of P. tabuliformis in Shanxi Province

    标记Locus FIS FIT FST Nm
    J20 0.330 8 0.362 8 0.047 8 4.980 7
    J9 -0.238 6 -0.226 0 0.010 1 24.383 3
    J42 0.100 0 0.146 4 0.051 6 4.590 5
    J48 -0.232 3 -0.157 1 0.061 0 3.848 0
    J29 0.492 9 0.502 5 0.019 0 12.935 5
    J50 0.000 5 0.004 0 0.003 5 70.972 6
    J10 -0.482 8 -0.469 0 0.009 3 26.725 3
    J12 0.014 5 0.055 6 0.041 6 5.755 2
    8个位点平均值Mean value of eight locus -0.056 5 -0.021 9 0.032 8 7.380 8
    4个位点平均值Mean value of four locus 0.053 8 0.102 9 0.051 9 4.570 3
    注:FST.杂合性基因多样度的比率;FIT.个体相对于总居群的固定指数;FIS.个体所在居群的固定指数;Nm.基因流。Notes: FST, ratio of heterozygosity gene diversity; FIT, fixed index of individual relative to total population; FIS, fixed index of the population in which the individual resides; Nm, gene flow.
    下载: 导出CSV

    表  7  油松种群遗传距离和地理距离

    Table  7.   Genetic distance and geographic distance among populations of P. tabuliformis in Shanxi Province

    山脉名称Mountain name GCS GDS THS TYS ZTS
    GCS —— 155.570 3 200.878 1 106.797 6 314.101 4
    GDS 0.006 7/0.009 8 —— 203.807 6 82.298 1 182.698 4
    THS 0.022 9/0.041 9 0.011 2/0.017 2 —— 134.989 2 221.989 3
    TYS 0.003 2/0.005 6 0.004 6/0.005 7 0.018 8/0.036 1 —— 208.215 3
    ZTS 0.026 2/0.051 2 0.021 2/0.038 9 0.040 7/0.074 4 0.025 4/0.049 3 ——
    注:表格左下方为油松种群遗传距离, 表格右上方为地理距离,km。遗传距离中“/”符号左方数据为8对引物分析结果,右方数据为4对引物分析结果。Notes: the bottom left of the table is the genetic distance of Pinus tabuliformis population. The upper right side of the table is geographic distance/km. The genetic distance data on the left of slash was analyzed by eight primers, and the genetic distance data on the right of slash was analyzed by four primers.
    下载: 导出CSV

    表  8  油松种群的遗传多样性参数与气象因子之间相关分析

    Table  8.   Correlation analysis on genetic diversity parameters and meteorological factors of P. tabuliformis populations

    指标Index Na Ne I Ho He
    8对引物
    Eight pair of primers
    4对引物
    Four pair of primers
    8对引物
    Eight pair of primers
    4对引物
    Four pair of primers
    8对引物
    Eight pair of primers
    4对引物
    Four pair of primers
    8对引物
    Eight pair of primers
    4对引物
    Four pair of primers
    8对引物
    Eight pair of primers
    4对引物
    Four pair of primers
    年均温Annual mean temperature -0.598 -0.662 -0.532 -0.565 -0.517 -0.581 -0.727 -0.790 -0.451 -0.532
    1月均温Average temperature in January -0.761 -0.801 -0.430 -0.432 -0.644 -0.711 -0.709 -0.822 -0.539 -0.651
    7月均温Average temperature in July -0.417 -0.511 -0.623 -0.671 -0.409 -0.457 -0.707 -0.710 -0.394 -0.430
    年均降水Annual mean precipitation -0.367 -0.535 0.040 -0.040 -0.392 -0.535 -0.155 -0.277 -0.322 -0.503
    年均降水与1月均温比值Ratio of annul mean precipitation and average temperature in January 0.780 0.922 0.470 0.459 0.776 0.875 0.722 0.831 0.693 0.830
    年均降水与7月均温比值Ratio of annual mean precipitation and average temperature in July -0.118 -0.180 0.340 0.367 -0.115 -0.194 0.210 0.109 -0.057 -0.178
    年均降水与年均温比值Ratio of annual mean precipitation and annual mean temperature 0.280 0.307 0.528 0.573 0.242 0.252 0.570 0.551 0.237 0.230
    下载: 导出CSV
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  • 收稿日期:  2018-02-14
  • 修回日期:  2018-07-03
  • 刊出日期:  2018-10-01

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